Consumers' health would be at risk unless food safety is safeguarded by food handlers in establishments. The study aimed to investigate food safety practices and its determinants using a cross-sectional study conducted on 420 food handlers in Ethiopia Ambo Town. Logistic regression analysis was adopted to analyze the data. A total of 415 food handlers participated with a response rate of 98.8 %. About 48.6% of the respondents had good food safety practices. The adjusted odd ratio (AOR) and confidence interval (CI) of having food safety training [AOR = 6.05, 95% CI: 3.49–10.49], medical check-ups [AOR = 2.07, 95% CI: 1.11–3.86], inspected by health professions [AOR = 2.26, 95% CI: 1.32–3.88], work experience [AOR = 4.71, 95% CI: 2.11–10.50], and good food safety knowledge [AOR = 2.25, 95% CI: 1.32–3.86] were significantly associated with food safety practices. Poor food safety practices were assessed in Ambo Town, Ethiopia.